PMID- 36461917
OWN - NLM
STAT- MEDLINE
DCOM- 20230214
LR  - 20231204
IS  - 1532-2548 (Electronic)
IS  - 0032-0889 (Print)
IS  - 0032-0889 (Linking)
VI  - 191
IP  - 2
DP  - 2023 Feb 12
TI  - Ascorbate peroxidase 1 allows monitoring of cytosolic accumulation of 
      effector-triggered reactive oxygen species using a luminol-based assay.
PG  - 1416-1434
LID - 10.1093/plphys/kiac551 [doi]
AB  - Biphasic production of reactive oxygen species (ROS) has been observed in plants 
      treated with avirulent bacterial strains. The first transient peak corresponds to 
      pattern-triggered immunity (PTI)-ROS, whereas the second long-lasting peak 
      corresponds to effector-triggered immunity (ETI)-ROS. PTI-ROS are produced in the 
      apoplast by plasma membrane-localized NADPH oxidases, and the recognition of an 
      avirulent effector increases the PTI-ROS regulatory module, leading to ETI-ROS 
      accumulation in the apoplast. However, how apoplastic ETI-ROS signaling is 
      relayed to the cytosol is still unknown. Here, we found that in the absence of 
      cytosolic ascorbate peroxidase 1 (APX1), the second phase of ETI-ROS accumulation 
      was undetectable in Arabidopsis (Arabidopsis thaliana) using luminol-based 
      assays. In addition to being a scavenger of cytosolic H2O2, we discovered that 
      APX1 served as a catalyst in this chemiluminescence ROS assay by employing 
      luminol as an electron donor. A horseradish peroxidase (HRP)-mimicking APX1 
      mutation (APX1W41F) further enhanced its catalytic activity toward luminol, 
      whereas an HRP-dead APX1 mutation (APX1R38H) reduced its luminol oxidation 
      activity. The cytosolic localization of APX1 implies that ETI-ROS might 
      accumulate in the cytosol. When ROS were detected using a fluorescent dye, green 
      fluorescence was observed in the cytosol 6 h after infiltration with an avirulent 
      bacterial strain. Collectively, these results indicate that ETI-ROS eventually 
      accumulate in the cytosol, and cytosolic APX1 catalyzes luminol oxidation and 
      allows monitoring of the kinetics of ETI-ROS in the cytosol. Our study provides 
      important insights into the spatial dynamics of ROS accumulation in plant 
      immunity.
CI  - (c) American Society of Plant Biologists 2022. All rights reserved. For 
      permissions, please e-mail: journals.permissions@oup.com.
FAU - Hong, Xiufang
AU  - Hong X
AUID- ORCID: 0000-0001-9929-6945
AD  - State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and 
      Safety of Agro-products, Institute of Biotechnology, Zhejiang University, 
      Hangzhou 310058, China.
FAU - Qi, Fan
AU  - Qi F
AUID- ORCID: 0000-0002-1963-1481
AD  - State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and 
      Safety of Agro-products, Institute of Biotechnology, Zhejiang University, 
      Hangzhou 310058, China.
FAU - Wang, Ran
AU  - Wang R
AD  - State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and 
      Safety of Agro-products, Institute of Biotechnology, Zhejiang University, 
      Hangzhou 310058, China.
FAU - Jia, Zhiyi
AU  - Jia Z
AUID- ORCID: 0000-0002-2153-8817
AD  - State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and 
      Safety of Agro-products, Institute of Biotechnology, Zhejiang University, 
      Hangzhou 310058, China.
FAU - Lin, Fucheng
AU  - Lin F
AUID- ORCID: 0000-0002-4127-8143
AD  - State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and 
      Safety of Agro-products, Institute of Biotechnology, Zhejiang University, 
      Hangzhou 310058, China.
FAU - Yuan, Minhang
AU  - Yuan M
AUID- ORCID: 0000-0001-6184-2998
AD  - National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in 
      Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese 
      Academy of Sciences, Shanghai 200032, China.
FAU - Xin, Xiu-Fang
AU  - Xin XF
AUID- ORCID: 0000-0002-4037-3528
AD  - National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in 
      Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese 
      Academy of Sciences, Shanghai 200032, China.
FAU - Liang, Yan
AU  - Liang Y
AUID- ORCID: 0000-0002-1248-8563
AD  - State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and 
      Safety of Agro-products, Institute of Biotechnology, Zhejiang University, 
      Hangzhou 310058, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Plant Physiol
JT  - Plant physiology
JID - 0401224
RN  - 0 (Reactive Oxygen Species)
RN  - EC 1.11.1.11 (Ascorbate Peroxidases)
RN  - 0 (Arabidopsis Proteins)
RN  - 5EXP385Q4F (Luminol)
RN  - BBX060AN9V (Hydrogen Peroxide)
SB  - IM
MH  - Reactive Oxygen Species
MH  - Ascorbate Peroxidases/genetics
MH  - *Arabidopsis Proteins/genetics
MH  - Luminol
MH  - Cytosol
MH  - Hydrogen Peroxide
MH  - *Arabidopsis/microbiology
PMC - PMC9922408
COIS- Conflict of interest statement. The authors declare that they have no conflict of 
      interest.
EDAT- 2022/12/04 06:00
MHDA- 2023/02/15 06:00
PMCR- 2023/12/03
CRDT- 2022/12/03 08:42
PHST- 2022/05/18 00:00 [received]
PHST- 2022/12/02 00:00 [accepted]
PHST- 2022/12/04 06:00 [pubmed]
PHST- 2023/02/15 06:00 [medline]
PHST- 2022/12/03 08:42 [entrez]
PHST- 2023/12/03 00:00 [pmc-release]
AID - 6869129 [pii]
AID - kiac551 [pii]
AID - 10.1093/plphys/kiac551 [doi]
PST - ppublish
SO  - Plant Physiol. 2023 Feb 12;191(2):1416-1434. doi: 10.1093/plphys/kiac551.